Short communication
A luciferase immunosorbent assay for quantitative detection of IgG antibodies against SARS-CoV-2 nucleoprotein

https://doi.org/10.1016/j.jviromet.2021.114141Get rights and content

Highlights

  • LISA uses the crude cell extracts directly rather than the purified viral protein, so it is a time-saving testing platform.

  • LISA measures the levels of antibody according to the luciferase light units and shows a wide linear range of quantitation.

  • LISA does not need species-specific secondary antibodies, so it is universal for detecting the specific antibody in a wide range of species.

Abstract

In this study, we developed and evaluated a luciferase immunosorbent assay (LISA) for quantitative detection of IgG antibody against SARS-CoV-2 nucleoprotein (NP). Anti-SARS-CoV-2 NP antibody in serum or plasma samples was captured by protein G-coated microtiter plate and detected using the crude cell lysates expressing Nanoluc luciferase (Nluc) enzyme fused with SARS-CoV-2 NP. After the addition of furimazine substrate, the levels of anti-SARS-CoV-2 NP IgG antibody were quantitatively measured as luciferase light units. As expected, SARS-CoV-2 NP showed cross-reactivity with the monoclonal antibodies against SARS-CoV NP, but not MERS-CoV NP-specific monoclonal antibodies or the monoclonal antibodies against SARS-CoV Spike protein. LISA for detecting murine monoclonal antibody against SARS-CoV NP showed a low limit of detection of 0.4 pg/μl and linear detection range from 0.4 pg/μl to 75 pg/μl. Furthermore, LISA had a sensitivity of 71 % when testing COVID-19 patients at the second week post onset and a specificity of 100 % when testing healthy blood donors.

Keywords

Luciferase immunosorbent assay
SARS-CoV-2
Semi-quantitative
Antibody detection

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